ZZ Domain of CBP: an Unusual Zinc Finger Fold in a Protein Interaction Module

Legge, Glen B.; Martinez-Yamout, Maria A.; Hambly, David M.; Trinh, Tam; Lee, Brian M.; Dyson, H. Jane; Wright, Peter E.

doi:10.1016/j.jmb.2004.08.087

Cited by 80 publications

(79 citation statements)

References 66 publications

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“…The TAZ2 domain of mouse CBP (residues 1764 -1855) and the p53 transactivation domain (residues 13-61) were expressed and purified as described (36,48). The pRb central pocket and adenovirus serotype 5 (Ad5 and Ad6) E1A constructs were expressed in Escherichia coli BL21 (DE3) cells; details are given in SI Text.…”

Section: Methodsmentioning

confidence: 99%

Structural basis for subversion of cellular control mechanisms by the adenoviral E1A oncoprotein

Ferreon

Martinez-Yamout²,

Dyson³

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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119

186

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The adenovirus early region 1A (E1A) oncoprotein mediates cell transformation by deregulating host cellular processes and activating viral gene expression by recruitment of cellular proteins that include cyclic-AMP response element binding (CREB) binding protein (CBP)/p300 and the retinoblastoma protein (pRb). While E1A is capable of independent interaction with CBP/p300 or pRb, simultaneous binding of both proteins is required for maximal biological activity. To obtain insights into the mechanism by which E1A hijacks the cellular transcription machinery by competing with essential transcription factors for binding to CBP/p300, we have determined the structure of the complex between the transcriptional adaptor zinc finger-2 (TAZ2) domain of CBP and the conserved region-1 (CR1) domain of E1A. The E1A CR1 domain is unstructured in the free state and upon binding folds into a local helical structure mediated by an extensive network of intermolecular hydrophobic contacts. By NMR titrations, we show that E1A efficiently competes with the N-terminal transactivation domain of p53 for binding to TAZ2 and that pRb interacts with E1A at 2 independent sites located in CR1 and CR2. We show that pRb and the CBP TAZ2 domain can bind simultaneously to the CR1 site of E1A to form a ternary complex and propose a structural model for the pRb:E1A:CBP complex on the basis of published x-ray data for homologous binary complexes. These observations reveal the molecular basis by which E1A inhibits p53-mediated transcriptional activation and provide a rationale for the efficiency of cellular transformation by the adenoviral E1A oncoprotein.CBP/p300 ͉ NMR ͉ protein structure ͉ transcriptional coactivator ͉ retinoblastoma protein T he adenovirus early region 1A (E1A) protein plays a central function in viral infection by activating genes required for viral replication and by deregulating the host cell cycle to force entry into S phase and repress differentiation (1-3). E1A deregulates the cell cycle through interactions with key cellular proteins, including the general transcriptional coactivators cyclic-AMP response element binding (CREB) binding protein (CBP) and p300 (4, 5) and the retinoblastoma protein (pRb) (6), which lead to epigenetic modifications and reprogramming of host cell transcriptional processes (7). Binding of E1A to pRb displaces the E2F transcription factors, thereby relieving E2F repression and resulting in premature S-phase entry and transcriptional activation of E2F-regulated genes (8-10). Association of E1A with CBP/p300 results in global hypoacetylation of K18 of histone H3 and may be linked to the ability of E1A to induce oncogenic transformation (11). E1A is a multifunctional protein (12) composed of 4 conserved regions (CR1-CR4, Fig. 1A). E1A uses both CR1 and CR2 for interaction with pRb (13,14). CR2 contains a characteristic pRb recognition motif, LX-CXE, which binds with high affinity to the B cyclin fold domain of the pRb pocket region (15, 16). The CR1 region of E1A binds at the interface of the A and B ...

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Section: Methodsmentioning

confidence: 99%

Structural basis for subversion of cellular control mechanisms by the adenoviral E1A oncoprotein

Ferreon

Martinez-Yamout²,

Dyson³

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

119

186

View full text Add to dashboard Cite

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“…Additionally, both RING/U-box domains, as well as both UPF1 RING-like modules, are similar in structure to the ZZ domain of CREB-binding protein (CBP), another twozinc binding module found in diverse proteins ( Fig. 3F; Legge et al 2004). The ZZ domain has been proposed to function in protein-protein interactions but is not well characterized.…”

Section: Zinc Coordination Within the Upf1 Ch-rich Domainmentioning

confidence: 96%

“…The correct folding of the UPF1 mutants that affected UPF2 binding (V204D, V206E, E182R + Y184D, F192E, V161E + R162E, V206E + V161E + R162E, Y125E) was confirmed by size-exclusion chromatography, which showed them to behave identically to the native protein (data not shown). Moreover these experiments (Legge et al 2004) (PDB entry 1TOT). The UPF1 RING-like module 1 and the ZZ domain superpose with an RMS deviation of 2.03 Å for 32 C a atoms (DALI Z-score of 3.6).…”

Section: Mutational Analysis Of Upf1-upf2 Interactionmentioning

confidence: 99%

Crystal structure of the UPF2-interacting domain of nonsense-mediated mRNA decay factor UPF1

Kadlec

Guilligay²,

Ravelli³

et al. 2006

RNA

109

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UPF1 is an essential eukaryotic RNA helicase that plays a key role in various mRNA degradation pathways, notably nonsensemediated mRNA decay (NMD). In combination with UPF2 and UPF3, it forms part of the surveillance complex that detects mRNAs containing premature stop codons and triggers their degradation in all organisms studied from yeast to human. We describe the 3 Å resolution crystal structure of the highly conserved cysteine-histidine-rich domain of human UPF1 and show that it is a unique combination of three zinc-binding motifs arranged into two tandem modules related to the RING-box and U-box domains of ubiquitin ligases. This UPF1 domain interacts with UPF2, and we identified by mutational analysis residues in two distinct conserved surface regions of UPF1 that mediate this interaction. UPF1 residues we identify as important for the interaction with UPF2 are not conserved in UPF1 homologs from certain unicellular parasites that also appear to lack UPF2 in their genomes.

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“…The ZZ domain consists of two anti-parallel ␤-sheets and a short ␣-helix coordinating two zinc ions as revealed by the first published ZZ domain structure originating from CBP (CREB-binding protein/p300) (11). There are ϳ20 ZZ domains (11,12) identified in the human genome, but a general biological function has not been assigned.…”

Section: Sumomentioning

confidence: 99%

Structural Analysis of a Complex between Small Ubiquitin-like Modifier 1 (SUMO1) and the ZZ Domain of CREB-binding Protein (CBP/p300) Reveals a New Interaction Surface on SUMO

Diehl

Akke

Bekker‐Jensen

et al. 2016

Journal of Biological Chemistry

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We have recently discovered that the ZZ zinc finger domain represents a novel small ubiquitin-like modifier (SUMO) binding motif. In this study we identify the binding epitopes in the ZZ domain of CBP (CREB-binding protein) and SUMO1 using NMR spectroscopy. The binding site on SUMO1 represents a unique epitope for SUMO interaction spatially opposite to that observed for canonical SUMO interaction motifs (SIMs). HAD-DOCK docking simulations using chemical shift perturbations and residual dipolar couplings was employed to obtain a structural model for the ZZ domain-SUMO1 complex. Isothermal titration calorimetry experiments support this model by showing that the mutation of key residues in the binding site abolishes binding and that SUMO1 can simultaneously and noncooperatively bind both the ZZ domain and a canonical SIM motif. The binding dynamics of SUMO1 was further characterized using 15 N Carr-Purcell-Meiboom-Gill (CPMG) relaxation dispersions, which define the off rates for the ZZ domain and SIM motif and show that the dynamic binding process has different characteristics for the two cases. Furthermore, in the absence of bound ligands SUMO1 transiently samples a high energy conformation, which might be involved in ligand binding. SUMO2 (small ubiquitin-like modifier) is a protein structurally homologous to ubiquitin that functions as a post-translational modifier involved in diverse cellular processes such as chromatin remodeling, ubiquitin E3 ligation, autophagy, cytoskeletal scaffolding, and DNA repair. Higher eukaryotes encode at least three different functional SUMO isoforms, in the human genome designated as SUMO1-3, which can conjugate to a number of target proteins (1-3). SUMO modified target proteins are recognized by SUMO interacting motifs (SIMs), the minimal consensus sequence, which is KXE, where consists of a large hydrophobic amino acid and where X can be any amino acid (4, 5). In addition to this canonical type of SIM, closely related variants such as inverted SIMs and phosphorylation-dependent SIMs have also been recently described (6 -8). The binding site for the canonical SIM is located in a groove between the ␣-helix and ␤-sheet in SUMO, where the SIM motif can bind either in a parallel or anti-parallel fashion (6).We recently showed that the ubiquitin ligase HERC2 can bind SUMO1 through its zinc finger ZZ domain (9), which therefore represents a new class of SUMO binding motifs. Isothermal titration calorimetry (ITC) showed that the ZZ domain binds to SUMO1 with M affinity in the same range as the canonical SIMs (10). The ZZ domain consists of two anti-parallel ␤-sheets and a short ␣-helix coordinating two zinc ions as revealed by the first published ZZ domain structure originating from CBP (CREB-binding protein/p300) (11). There are ϳ20 ZZ domains (11, 12) identified in the human genome, but a general biological function has not been assigned.Here we present the complex between SUMO1 and the ZZ domain from CBP, where we map the interaction interface using NMR spectroscopy. The structure ...

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ZZ Domain of CBP: an Unusual Zinc Finger Fold in a Protein Interaction Module

Cited by 80 publications

References 66 publications

Structural basis for subversion of cellular control mechanisms by the adenoviral E1A oncoprotein

Structural basis for subversion of cellular control mechanisms by the adenoviral E1A oncoprotein

Crystal structure of the UPF2-interacting domain of nonsense-mediated mRNA decay factor UPF1

Structural Analysis of a Complex between Small Ubiquitin-like Modifier 1 (SUMO1) and the ZZ Domain of CREB-binding Protein (CBP/p300) Reveals a New Interaction Surface on SUMO

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